Nori Equine ApoAI ELISA Kit
$461.00 – $832.00
This ELISA kit is for quantification of ApoAI in horse. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for ApoAI has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any ApoAI present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for ApoAI is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of ApoAI bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for ApoAI: Apolipoprotein A-I, ApoA1,
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Equine ApoAI ELISA Kit Summary
Alternative names for ApoAI: Apolipoprotein A-I, ApoA1,
Alternative name for equine: Horse
| Assay Type | Solid Phase Sandwich ELISA |
| Format | 96-well Microplate or 96-Well Strip Microplate |
| Method of Detection | Colorimetric |
| Number of Targets Detected | 1 |
| Target Antigen Accession Number | F6Z2L5 |
| Assay Length | 3 hours |
| Quantitative/Semiquantitative | Quantitative |
| Sample Type | Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL, |
| Recommended Sample Dilution (Plasma/Serum) | No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ |
| Sensitivity | 300 pg/mL |
| Detection Range | 1.56-100 ng/mL |
| Specificity | Equine ApoAI |
| Cross-Reactivity | < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested. |
| Interference | No significant interference observed with available related molecules |
| Storage/Stability | 4 ºC for up to 6 months |
| Usage | For Laboratory Research Use Only. Not for diagnostic or therapeutic use. |
| Additional Notes | The kit allows for use in multiple experiments. |
Standard Curve
Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer
Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.
Background:
Apolipoprotein A-I is the major protein component of high-density lipoprotein (HDL) in plasma. Chylomicrons secreted from the intestinal enterocyte also contain ApoA1 but it is quickly transferred to HDL in the bloodstream (1). The protein promotes cholesterol efflux from tissues to the liver for excretion. It is a cofactor for lecithin cholesterol acyltransferase (LCAT) which is responsible for the formation of most plasma cholesteryl esters. ApoA-I was also isolated as a prostacyclin (PGI2) stabilizing factor, and thus may have an anticlotting effect (2). Defects in the gene encoding it are associated with HDL deficiencies, including Tangier disease, and with systemic non-neuropathic amyloidosis (3). A G/A polymorphism in the promoter of the ApoA-I gene has been associated with the age at which patients presented with Alzheimer disease (4). Protection from Alzheimer disease by ApoA1 may rely on a synergistic interaction with alpha-tocopherol (5). Amyloid deposited in the knee following surgery consists largely of ApoA-I secreted from chondrocytes (cartilage cells) (6). A wide variety of amyloidosis symptoms are associated with rare ApoA-I mutants. ApoA-I binds to lipopolysaccharide or endotoxin, and has a major role in the anti-endotoxin function of HDL.
References
- Wasan KM, et al. (2008). Nature Reviews Drug Discovery 7 (1): 84
- Yui Y, et al. (1988). J. Clin. Invest. 82 (3): 803–7.
- Entrez Gene: APOA1 apolipoprotein A-I.
- Vollbach H, et al. (2005). Ann. Neurol. 58 (3): 436
- Maezawa I, at al. (2004). J. Neurochem. 91 (6): 1312
- Solomon A, et al. (2006). Arthritis Rheum. 54 (11): 3545.
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